来自脆双核阿米巴的锥虫硫醇还原酶的初步晶体学分析。

Preliminary crystallographic analysis of trypanothione reductase from Crithidia fasciculata.

作者信息

Kuriyan J, Wong L, Guenther B D, Murgolo N J, Cerami A, Henderson G B

机构信息

Laboratory of Molecular Biophysics, Rockefeller University, New York, NY 10021.

出版信息

J Mol Biol. 1990 Oct 5;215(3):335-7. doi: 10.1016/s0022-2836(05)80353-5.

DOI:10.1016/s0022-2836(05)80353-5

PMID:2231707

Abstract

Trypanothione reductase, a flavoprotein disulfide reductase specific to trypanosomatid parasites, has been crystallized by vapor diffusion of a protein solution (10 mg/ml) against 22% polyethylene glycol (average Mr 8000) containing 100 mM-ammonium sulfate. Crystals of a size suitable for structure determination by X-ray diffraction have been obtained by seeding protein solutions with smaller crystals. The space-group is P21 (a = 60.9 A, b = 161.8 A, c = 58.4 A, beta = 99.1 degrees). The molecular mass and volume of the unit cell suggest that there is a dimer of the enzyme in the asymmetric unit, and this is confirmed by self-rotation functions calculated using data to 4.5 A resolution. The crystals diffract to beyond 3 A resolution. Crystals of another P21 form (a = 91.3 A, b = 114.4 A, c = 92.0 A, beta = 141.3 degrees) are observed to grow under similar conditions.

摘要

锥虫硫醇还原酶是一种锥虫科寄生虫特有的黄素蛋白二硫化物还原酶，它是通过将蛋白质溶液（10 mg/ml）与含有100 mM硫酸铵的22%聚乙二醇（平均分子量8000）进行气相扩散而结晶的。通过用较小的晶体对蛋白质溶液进行接种，已获得了适合通过X射线衍射进行结构测定的晶体。空间群为P21（a = 60.9 Å，b = 161.8 Å，c = 58.4 Å，β = 99.1°）。晶胞的分子量和体积表明不对称单元中有该酶的二聚体，这通过使用分辨率为4.5 Å的数据计算的自旋转函数得到证实。这些晶体的衍射分辨率超过3 Å。在类似条件下观察到另一种P21形式的晶体（a = 91.3 Å，b = 114.4 Å，c = 92.0 Å，β = 141.3°）生长。